NOTE: End date changed to 12/31/2024 per PI (Ed., 8/25/23)

NOTE: End date changed to 09/30/2023 per T. Sirmons/JSC (Ed., 5/16/23)

NOTE: End date changed to 12/31/2022 per PI (Ed., 4/23/21)

NOTE: Extended to 7/31/2021 per PI (Ed., 8/6/19)

NOTE: Extended to 3/30/2020 per PI (Ed., 2/7/19)

NOTE: End date changed to 9/30/2019 due to revised research plan with Ground and Flight work, per HRP (Ed., 8/24/16)

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

There is a low -- but real -- incidence of infectious disease among crewmembers during spaceflight. Compared to ground controls, the incidence of latent herpesvirus reactivation is higher than expected. In addition, there is a consistent incidence of allergy and hypersensitivity symptoms; antihistamines remain the second-most prescribed medication onboard the International Space Station (ISS). Furthermore, data from flight studies suggest that during spaceflight astronauts exhibit persistent, low-level inflammation. All of these morbidities may be manifestations of altered immune function. However, the breadth of the perturbations throughout the human immune system, as well as their persistence during long-duration space missions, are unknown. If spaceflight impairs stably multiple aspects of the immune system, then it may confer a serious clinical risk to crewmembers for exploration-class missions.

We hypothesize that while aspects of adaptive immune function are depressed during spaceflight, aspects of humoral or innate immune function may be unaltered or even sensitized. This would explain the observed reactivation of latent herpesviruses in astronauts, and also the elevated incidence of skin rashes and hypersensitivity reactions during International Space Station (ISS) missions. If the hypothesis is validated, it would be an important consideration for any future immunology countermeasures. For example, one would not give an immune ‘booster’ to address T cell function in a crewmember, if it might potentially worsen on-orbit skin rashes or allergy symptoms. We further hypothesize that there is a widely disparate post-landing recovery for various aspects of immune dysregulation following flight. Previous data have demonstrated that, surprisingly, ISS astronauts maintain shedding of latent herpesviruses at least to R+30. This study will fully characterize all relevant immune dysregulation through a post-mission recovery.

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters.

Specific Aim 1: To define longitudinally the functional capacity of adaptive and innate immune cells. For T cells, our endpoints are activation status and cytokine production; for Natural Killer (NK) cells, the endpoint is cytotoxic (killing) ability.

Specific Aim 2: To determine the effect of spaceflight on a suite of soluble biomarkers that communicate in-vivo immune and physiological competence. From plasma, saliva, and urine we will quantify protein biomarkers of stress, inflammation, antimicrobial activity, and latent viral reactivation. Several solicited parameters will augment this aim, including proteomics and/or genomics.

Specific Aim 3: To relate the immune profiles of astronauts to their clinical status, as well as to their habitual behavior in space: sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA). The results will inform NASA’s scientific and operational communities about the influence on immunity of spaceflight-specific activity, factors we may modulate as part of an immune countermeasures strategy.

The conclusions from this study will determine whether or not immune countermeasures will be necessary for exploration space missions. These data will define an immune surveillance strategy, comprehensive in its scope to encompass innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be critical to validate candidate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

Additionally, we had enrolled one astronaut assigned to launch from a vehicle that had experienced several technical delays; therefore, that subject was also dropped from the study after collecting one preflight time-point. Although we achieved our target enrollment, an insurance astronaut was recruited, along with a suitable ground control subject; we enrolled this study subject at the beginning of 2021.

We have completed sampling for the end total of 11 astronauts and their ground control subjects for all mission time-points. The specimens from all the time-points we collected have been processed: samples of whole blood, plasma, and peripheral blood mononuclear cells (PBMC) were partitioned and sent to the external collaborators; plasma was isolated and cryopreserved for cytokine analysis; immune cells were assayed for function and phenotype via flow cytometry; supernatants from 48hr stimulation cultures were cryopreserved for measurement of cytokine content. This closed out the sampling phase of the study.

We dispatched all respective samples to our external collaborators at the time of initial receipt, after initial processing, or in bulk shipments at strategic intervals. All samples have been successfully transferred to each external collaborator and their batch analyses of the specimens is completed. Each collaborator is writing up their final longitudinal analysis report to send to NASA individually.

We completed analysis of the flow cytometry data at every time-point for every crew and ground control subject, and we are currently synthesizing them via longitudinal analysis. The virology lab processed the saliva and urine specimens at every time-point collected for all crew and ground control subjects, performed the experiments to quantify stress hormones and latent virus reactivation, and recorded the data. Virology is synthesizing data via longitudinal analysis.

A cross-analysis combined final report will be submitted later.

NOTE: End date changed to 12/31/2024 per PI (Ed., 8/25/23)

NOTE: End date changed to 09/30/2023 per T. Sirmons/JSC (Ed., 5/16/23)

NOTE: End date changed to 12/31/2022 per PI (Ed., 4/23/21)

NOTE: Extended to 7/31/2021 per PI (Ed., 8/6/19)

NOTE: Extended to 3/30/2020 per PI (Ed., 2/7/19)

NOTE: End date changed to 9/30/2019 due to revised research plan with Ground and Flight work, per HRP (Ed., 8/24/16)

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

There is a low -- but real -- incidence of infectious disease among crewmembers during spaceflight. Compared to ground controls, the incidence of latent herpesvirus reactivation is higher than expected. In addition, there is a consistent incidence of allergy and hypersensitivity symptoms; antihistamines remain the second-most prescribed medication onboard the International Space Station (ISS). Furthermore, data from flight studies suggest that during spaceflight astronauts exhibit persistent, low-level inflammation. All of these morbidities may be manifestations of altered immune function. However, the breadth of the perturbations throughout the human immune system, as well as their persistence during long-duration space missions, are unknown. If spaceflight impairs stably multiple aspects of the immune system, then it may confer a serious clinical risk to crewmembers for exploration-class missions.

We hypothesize that while aspects of adaptive immune function are depressed during spaceflight, aspects of humoral or innate immune function may be unaltered or even sensitized. This would explain the observed reactivation of latent herpesviruses in astronauts, and also the elevated incidence of skin rashes and hypersensitivity reactions during International Space Station (ISS) missions. If the hypothesis is validated, it would be an important consideration for any future immunology countermeasures. For example, one would not give an immune ‘booster’ to address T cell function in a crewmember, if it might potentially worsen on-orbit skin rashes or allergy symptoms. We further hypothesize that there is a widely disparate post-landing recovery for various aspects of immune dysregulation following flight. Previous data have demonstrated that, surprisingly, ISS astronauts maintain shedding of latent herpesviruses at least to R+30. This study will fully characterize all relevant immune dysregulation through a post-mission recovery.

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters.

Specific Aim 1: To define longitudinally the functional capacity of adaptive and innate immune cells. For T cells, our endpoints are activation status and cytokine production; for Natural Killer (NK) cells, the endpoint is cytotoxic (killing) ability.

Specific Aim 2: To determine the effect of spaceflight on a suite of soluble biomarkers that communicate in-vivo immune and physiological competence. From plasma, saliva, and urine we will quantify protein biomarkers of stress, inflammation, antimicrobial activity, and latent viral reactivation. Several solicited parameters will augment this aim, including proteomics and/or genomics.

Specific Aim 3: To relate the immune profiles of astronauts to their clinical status, as well as to their habitual behavior in space: sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA). The results will inform NASA’s scientific and operational communities about the influence on immunity of spaceflight-specific activity, factors we may modulate as part of an immune countermeasures strategy.

The conclusions from this study will determine whether or not immune countermeasures will be necessary for exploration space missions. These data will define an immune surveillance strategy, comprehensive in its scope to encompass innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be critical to validate candidate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

To date, 11 astronauts and their ground control subjects have completed their sampling regimen of all mission time points. The specimens from all the time-points we collected have been processed: samples of whole blood, plasma, and peripheral blood mononuclear cells (PBMC) were partitioned and sent to the external collaborators; plasma was isolated and cryopreserved for cytokine analysis; immune cells were assayed for function and phenotype via flow cytometry; supernatants from 48hr stimulation cultures were cryopreserved for measurement of cytokine content. This closes out the sampling phase of the study. We analyzed the flow cytometry data at every time-point, and we are currently synthesizing them via longitudinal analysis.

We dispatched samples to our external collaborators at the time of initial receipt, after initial processing, or in bulk shipments at strategic intervals. All samples have been collected for all crew and matched control for each time point and have been successfully transferred to each external collaborator. They are currently performing batch analyses of the specimens and will report their findings to NASA individually. The virology lab processed the saliva and urine specimens at every time-point collected, performed the experiments to quantify stress hormones and latent virus reactivation, and recorded the data, but they have not synthesized them via longitudinal analysis yet.

A cross-analysis combined final report will be submitted later.

NOTE: End date changed to 12/31/2022 per PI (Ed., 4/23/21)

NOTE: Extended to 7/31/2021 per PI (Ed., 8/6/19)

NOTE: Extended to 3/30/2020 per PI (Ed., 2/7/19)

NOTE: End date changed to 9/30/2019 due to revised research plan with Ground and Flight work, per HRP (Ed., 8/24/16)

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

The Functional Immune project is a multi-disciplinary flight study whose purpose is to define comprehensively the immune system’s response to long-duration spaceflight.

There is a low -- but real -- incidence of infectious disease among crewmembers during spaceflight. Compared to ground controls, the incidence of latent herpesvirus reactivation is higher than expected. In addition, there is a consistent incidence of allergy and hypersensitivity symptoms; antihistamines remain the second-most prescribed medication onboard the International Space Station (ISS). Furthermore, data from flight studies suggest that during spaceflight astronauts exhibit persistent, low-level inflammation. All of these morbidities may be manifestations of altered immune function. However, the breadth of the perturbations throughout the human immune system, as well as their persistence during long-duration space missions, are unknown. If spaceflight impairs stably multiple aspects of the immune system, then it may confer a serious clinical risk to crewmembers for exploration-class missions.

We hypothesize that while aspects of adaptive immune function are depressed during spaceflight, aspects of humoral or innate immune function may be unaltered or even sensitized. This would explain the observed reactivation of latent herpesviruses in astronauts, and also the elevated incidence of skin rashes and hypersensitivity reactions during International Space Station (ISS) missions. If the hypothesis is validated, it would be an important consideration for any future immunology countermeasures. For example, one would not give an immune ‘booster’ to address T cell function in a crewmember, if it might potentially worsen on-orbit skin rashes or allergy symptoms. We further hypothesize that there is a widely disparate post-landing recovery for various aspects of immune dysregulation following flight. Previous data have demonstrated that, surprisingly, ISS astronauts maintain shedding of latent herpesviruses at least to R+30. This study will fully characterize all relevant immune dysregulation through a post-mission recovery.

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters.

Specific Aim 1: To define longitudinally the functional capacity of adaptive and innate immune cells. For T cells, our endpoints are activation status and cytokine production; for Natural Killer (NK) cells, the endpoint is cytotoxic (killing) ability.

Specific Aim 2: To determine the effect of spaceflight on a suite of soluble biomarkers that communicate in-vivo immune and physiological competence. From plasma, saliva, and urine we will quantify protein biomarkers of stress, inflammation, antimicrobial activity, and latent viral reactivation. Several solicited parameters will augment this aim, including proteomics and/or genomics.

Specific Aim 3: To relate the immune profiles of astronauts to their clinical status, as well as to their habitual behavior in space: sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA). The results will inform NASA’s scientific and operational communities about the influence on immunity of spaceflight-specific activity, factors we may modulate as part of an immune countermeasures strategy.

The conclusions from this study will determine whether or not immune countermeasures will be necessary for exploration space missions. These data will define an immune surveillance strategy, comprehensive in its scope to encompass innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be critical to validate candidate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

We powered the Functional Immune project to investigate 10 astronauts and 10 gender- and age- matched control subjects. Enrollment for this study began in 2016, and the first sampling time-point occurred in August of that year. To date, we have enrolled 12 astronauts and 10 ground control subjects. Unfortunately, due to the delay in launching Expedition 58, the altered sampling timeframe for the inflight time-points did not conform to the experimental constraints of the study. Thus, we dropped from the study two of the enrolled astronauts who are aboard the Expedition 58 mission. Although we achieved our target enrollment, an insurance astronaut will be recruited, along with a suitable ground control subject; we expect to enroll this study subject towards the end of 2020.

To date, ten astronauts and their ground control subjects have completed their sampling regimen of seven time points. The specimens from all the time-points we collected have been processed: samples of whole blood, plasma, and peripheral blood mononuclear cell (PBMC) were partitioned and sent to the external collaborators; plasma was isolated and cryopreserved for cytokine analysis; immune cells were assayed for function and phenotype via flow cytometry; supernatants from 48 hr stimulation cultures were cryopreserved for measurement of cytokine content. We analyzed the flow cytometry data at every time-point, but we have not synthesized them via longitudinal analysis yet.

The virology lab processed the saliva and urine specimens at every time-point collected, performed the experiments to quantify stress hormones and latent virus reactivation, and recorded the data, but they have not synthesized them via longitudinal analysis yet.

We dispatch samples to our external collaborators at the time of initial receipt, after initial processing, or in bulk shipments at strategic intervals. They will perform batch analyses of the specimens and report their findings to NASA individually.

NOTE: Extended to 3/30/2020 per PI (Ed., 2/7/19)

NOTE: End date changed to 9/30/2019 due to revised research plan with Ground and Flight work, per HRP (Ed., 8/24/16)

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

The Functional Immune project is a multi-disciplinary flight study whose purpose is to define comprehensively the immune system’s response to long-duration spaceflight.

There is a low -- but real -- incidence of infectious disease among crewmembers during spaceflight. Compared to ground controls, the incidence of latent herpesvirus reactivation is higher than expected. In addition, there is a consistent incidence of allergy and hypersensitivity symptoms; antihistamines remain the second-most prescribed medication onboard the International Space Station (ISS). Furthermore, data from flight studies suggest that during spaceflight astronauts exhibit persistent, low-level inflammation. All of these morbidities may be manifestations of altered immune function. However, the breadth of the perturbations throughout the human immune system, as well as their persistence during long-duration space missions, are unknown. If spaceflight impairs stably multiple aspects of the immune system, then it may confer a serious clinical risk to crewmembers for exploration-class missions.

We hypothesize that while aspects of adaptive immune function are depressed during spaceflight, aspects of humoral or innate immune function may be unaltered or even sensitized. This would explain the observed reactivation of latent herpesviruses in astronauts, and also the elevated incidence of skin rashes and hypersensitivity reactions during International Space Station (ISS) missions. If the hypothesis is validated, it would be an important consideration for any future immunology countermeasures. For example, one would not give an immune ‘booster’ to address T cell function in a crewmember, if it might potentially worsen on-orbit skin rashes or allergy symptoms. We further hypothesize that there is a widely disparate post-landing recovery for various aspects of immune dysregulation following flight. Previous data have demonstrated that, surprisingly, ISS astronauts maintain shedding of latent herpesviruses at least to R+30. This study will fully characterize all relevant immune dysregulation through a post-mission recovery.

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters.

Specific Aim 1: To define longitudinally the functional capacity of adaptive and innate immune cells. For T cells, our endpoints are activation status and cytokine production; for Natural Killer (NK) cells, the endpoint is cytotoxic (killing) ability.

Specific Aim 2: To determine the effect of spaceflight on a suite of soluble biomarkers that communicate in-vivo immune and physiological competence. From plasma, saliva, and urine we will quantify protein biomarkers of stress, inflammation, antimicrobial activity, and latent viral reactivation. Several solicited parameters will augment this aim, including proteomics and/or genomics.

Specific Aim 3: To relate the immune profiles of astronauts to their clinical status, as well as to their habitual behavior in space: sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA). The results will inform NASA’s scientific and operational communities about the influence on immunity of spaceflight-specific activity, factors we may modulate as part of an immune countermeasures strategy.

The conclusions from this study will determine whether or not immune countermeasures will be necessary for exploration space missions. These data will define an immune surveillance strategy, comprehensive in its scope to encompass innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be critical to validate candidate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

The Functional Immune project benefits from a longitudinal sampling architecture, similar to the one that enabled the success of the Integrated Immune flight study previously. At every sampling time-point, we collect blood (ambient, live), saliva, and urine samples before, during, and following spaceflight. We powered the Functional Immune project to investigate 10 astronauts and 10 gender- and age- matched control subjects. Enrollment for this study began in 2016, and the first sampling time-point occurred in August of that year. To date, we have enrolled 11 astronauts and 10 ground control subjects. Unfortunately, due to the delay in launching the 57S expedition, the altered sampling timeframe for the inflight time-points does not conform to the experimental constraints of the study. Thus, we must drop from the study two of our enrolled astronauts who are aboard the 57S mission. We have replaced one of the crewmembers already, and recycled the associated control subject, since the gender and age are the same. Thus, we have nine crewmembers and nine ground control subjects towards our target of ten for each group. We expect to enroll the tenth study subject towards the end of 2019, and complete the sample collections during 2020.

To date, six astronauts and their ground control subjects have completed their sampling regimen, seven time points. For two other crewmembers and their controls there remains only the last time-point outstanding. The ninth crewmember and associated control completed their pre-launch sampling time-points recently. The specimens from all the time-points we collected have been processed: samples of whole blood, plasma, and peripheral blood mononuclear cell (PBMC) were partitioned and sent to the external collaborators; plasma was isolated and cryopreserved for future cytokine analysis; immune cells were assayed for function and phenotype via flow cytometry; supernatants from 48 hr stimulation cultures were cryopreserved for future measurement of cytokine content. We analyzed the flow cytometry data at every time-point, but we have not synthesized them via longitudinal analysis.

The virology lab processed the saliva and urine specimens at every time-point collected, performed the experiments to quantify stress hormones and latent virus reactivation, and recorded the data, but they have not synthesized them via longitudinal analysis. For the Microgen Laboratories work, PBMC samples have been cryopreserved; the samples will be batch-processed once all the time-points have been collected. Thus, we expect to receive data towards the end of 2019. Analysis of stimulated T cells through the use of confocal and electron microscopy from the completed subjects is also planned to begin soon.

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

This new task is proposed as a directed/solicited study that will engage multiple external and/or international collaborators. Specific rational for the directed portion of the study is as follows:

Validated assays - requirement for technical homology: Some of the same measurements previously demonstrated to be dysregulated during long-duration spaceflight should be continued during the new study. It is important to correlate demonstrated areas of dysregulation in adaptive or viral parameters with new areas of investigation in the same individual subjects. Otherwise the result is separate data sets among which it is impossible to perform correlative analysis to determine mechanistic interactions. Many of these assays, as implemented during the Integrated Immune flight study, were developed and fully validated (and published) at Johnson Space Center (JSC). To solicit for assays would introduce additional and unnecessary cost and delay, as well as invite technical differences that could have a negative effect on data correlations.

Validated sample processing protocols: For implementation of Integrated Immune, it was necessary to create and validate sample processing protocols (sample stability, storage conditions, methods to stain/fix/preserve, elimination of autofluorescence, etc.). In most cases, these requirements are unnecessary for terrestrial laboratories that do not deal with operational constraints. Terrestrial laboratories do not routinely analyze, nor do they even accept, aged samples. For example, sample preservation (cell, nucleic acid, culture products) must be stabilized to support IP participation.

Requirement for established collaborations: We anticipate that validate assays from international partner flight studies would be necessary for the success of this proposal. The NASA JSC investigators have established both flight and ground collaborations with other US and International Partner immunologists. Rather than repeat flight studies sequentially, it is desirable to collaborate for the planned studies, so that all relevant in-flight assays may be performed. Examples include assays from the Salivary Markers, Immuno, MoCISS flight studies, as well as the CHOICE and NEEMO (NASA Extreme Environment Mission Operations project) analog studies.

Implementation Aspects: For a complicated medical study with unique sampling constraints, the success of the Integrated Immune flight study has demonstrated the advantages of an in-house investigator component for study success. This is relevant to crew ICB, crew training, sample processing and distribution, and central coordination of sampling, sample sharing, maintenance of banked samples, sample storage for batch analysis, facilitation of data coordination for interpretative purposes, data coordination for report generation, interdisciplinary aspects (established linkages with Nutrition, Bone, Cardio disciplines, etc.), and interface with JSC flight surgeons, JSC data archives. Examples include Integrated Immune, where immune data are being coordinated with crew sleep/wake cycles and circadian misalignment, and the planning for the Salivary Markers flight study.

Operational constraints-- Requirement for integrated and Immediate Sample Processing: The solicited aspects for this study will essentially be a very complicated sample sharing activity. All samples, including in-flight samples, must be processed at JSC, as JSC is the location where all BDC samples are collected, and where all in-flight samples return. In-flight samples will be at the end of their viability lifespan, by the time they are collected on-orbit, through hatch closure, deorbit, Soyuz return, and direct aircraft return to JSC. At this time, sample processing must occur without delay. Often, due to short (or otherwise sub optimal samples) real time judgment calls must be made to allow the maximal scientific return. We propose that to ensure mission success, the experienced JSC (directed) investigator team carry the integration responsibility for the entire team. The JSC staff has extensive experience regarding the integration required to support multiple immune investigators. For Integrated Immune, a study that also consisted of in-house and external collaborators (Mercer University, Microgen Laboratories, etc.), our suggested processing/implementation design worked well. For human subject samples, sampling volumes must be minimal, yet maximal assays must be generated on that product. JSC staff has processed ESA (European Space Agency) samples for both the NEEMO and Antarctica studies, and are knowledgeable regarding successfully integrating multiple IP studies on minimal sample. Outside investigators do not routinely work within this constraints.

Cost and efficiency: Given the above requirements for assays, integration, sample processing, and that these unique and validated assays and sample processing protocols are in-place at JSC, it would introduce significant additional cost to ‘train’ an external lab perform these assays to the same fidelity.

TASK DESCRIPTION NOVEMBER 2014: Some aspects of adaptive cellular immunity have been characterized during spaceflight, while many other areas of immunity (humoral, innate, cell specific functional, etc.) have not. The objective of this project is to characterize multiple diverse facets of immunoregulation during long-duration spaceflight. We hypothesize that while aspects of adaptive immune function are depressed during spaceflight, aspects of humoral or innate immune function may be unaltered or even sensitized. This would explain the observed reactivation of latent herpesviruses in astronauts, and also the elevated incidence of skin rashes and hypersensitivity reactions during International Space Station (ISS) missions. If the hypothesis is validated, it would be an important consideration for any future immunology countermeasures. For example, one would not give an immune ‘booster’ to address T cell function in a crewmember, if it might potentially worsen on-orbit skin rashes or allergy symptoms. We further hypothesize that there is a widely disparate post-landing recovery for various aspects of immune dysregulation following flight. Previous data have demonstrated that, surprisingly, ISS astronauts maintain shedding of latent herpesviruses at least to R+30. This study will fully characterize all relevant immune dysregulation through a post-mission recovery.

Primary Aims:

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in Astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters. Specific scientific aims are as follows:

1. Longitudinally examine the effect of spaceflight on previously uninvestigated aspects of immunobiology including leukocyte distribution, various aspects of innate cellular function. Examined concurrently will be several previously validated assays of adaptive cellular distribution and function, to correlate within crewmembers both innate and adaptive immune dysregulation.

2. Examine the relationship in astronauts between immune function and various markers of in-vivo immune-physiological status, including plasma, salivary and urinary markers of stress, antimicrobial activity, and latent viral reactivation. Various solicited parameters are planned to augment this specific aim, including proteomics and/or genomics.

Secondary Aims

3. Correlate findings of immune status with astronaut environmental, human, and stress factors such as sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA), etc. This correlative work should allow conclusions regarding environmental factors, which may potentially be modulated, on immune status.

4. Incorporate a final data analysis into specific conclusions regarding the immunobiology of spaceflight and conclusions regarding the necessity, lack thereof, or targeted aspects of, immune countermeasures for spaceflight. Develop a refined monitoring strategy encompassing innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be appropriate to validate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

NOTE: Title change to "Functional Immune Alterations, Latent Herpesvirus Reactivation, Physiological Stress, and Clinical Incidence Onboard the International Space Station" per original proposal to HRP, per L. Milstead/HRP. Title on original information provided was "Innate and Adaptive Immune Function during Long-duration Spaceflight" (Ed., 8/23/16)

Primary Aims:

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in Astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters. Specific scientific aims are as follows:

1. Longitudinally examine the effect of spaceflight on previously uninvestigated aspects of immunobiology including leukocyte distribution, various aspects of innate cellular function. Examined concurrently will be several previously validated assays of adaptive cellular distribution and function, to correlate within crewmembers both innate and adaptive immune dysregulation.

2. Examine the relationship in astronauts between immune function and various markers of in-vivo immune-physiological status, including plasma, salivary and urinary markers of stress, antimicrobial activity, and latent viral reactivation. Various solicited parameters are planned to augment this specific aim, including proteomics and/or genomics.

Secondary Aims

3. Correlate findings of immune status with astronaut environmental, human, and stress factors such as sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA), etc. This correlative work should allow conclusions regarding environmental factors, which may potentially be modulated, on immune status.

4. Incorporate a final data analysis into specific conclusions regarding the immunobiology of spaceflight and conclusions regarding the necessity, lack thereof, or targeted aspects of, immune countermeasures for spaceflight. Develop a refined monitoring strategy encompassing innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be appropriate to validate countermeasures.

Rationale Feb. 2015: This research is directed because it contains highly constrained research. In order to determine exact times of flight days and for the exact times in which the spacecraft cabin is depressed to 10.2 psia (thus creating the hypoxic condition), a researcher needs to have access to the Archive Data Retrieval (ADRIFT) subprogram inside of the Java Mission Evaluation Workstation System (JMEWS) which is only available on site here at Johnson Space Center, and requires proper clearance.

Aims:

The primary purpose of the study is to determine both acute and chronic alterations in crewmember immunobiology (both innate + adaptive parameters) in conjunction with relevant parameters from other disciplines (nutritional, radiation, virology, host-pathogen, stress, etc.). Parameters will be examined longitudinally in Astronauts before, during, and following spaceflight. Post-flight assessments will be extended to determine the timecourse for full recovery of any dysregulated parameters. Specific scientific aims are as follows:

1. Longitudinally examine the effect of spaceflight on previously uninvestigated aspects of immunobiology including leukocyte distribution, various aspects of innate cellular function. Examined concurrently will be several previously validated assays of adaptive cellular distribution and function, to correlate within crewmembers both innate and adaptive immune dysregulation.

2. Examine the relationship in astronauts between immune function and various markers of in-vivo immune-physiological status, including plasma, salivary and urinary markers of stress, antimicrobial activity, and latent viral reactivation. Various solicited parameters are planned to augment this specific aim, including proteomics and/or genomics.

3. Correlate findings of immune status with astronaut environmental, human, and stress factors such as sleep/wake data, crew work schedules, surveys of in-flight symptomology and/or medication use (voluntary), vehicle docking/undocking, extravehicular activity (EVA), etc. This correlative work should allow conclusions regarding environmental factors, which may potentially be modulated, on immune status.

4. Incorporate a final data analysis into specific conclusions regarding the immunobiology of spaceflight and conclusions regarding the necessity, lack thereof, or targeted aspects of, immune countermeasures for spaceflight. Develop a refined monitoring strategy encompassing innate immunity, adaptive immunity, and relevant parameters from other disciplines, which will be appropriate to validate countermeasures.